Highly multiplexed spectral FLIM via physics informed data analysis.

Spectral fluorescence lifetime imaging (S-FLIM) allows for the simultaneous deconvolution of signal from multiple fluorophore species by leveraging both spectral and lifetime information. However, existing analyses still face multiple difficulties in decoding information collected from typical S-FLIM experiments. These include: using information from pre-calibrated spectra in environments that may differ from the cellular context in which S-FLIM experiments are performed; limitations in the ability to deconvolute species due to overlapping spectra; high photon budget requirements, typically about a hundred photons per pixel per species.

Amino acid osmolytes disrupting intradomain interactions reduce the amyloidogenicity of α-synuclein: studies with charged l-amino acids and their derivatives.

Aggregation of α-synuclein (α-syn) is a hallmark of Parkinson's and dementia with Lewy bodies pathogenesis. The high plasticity and lack of stable tertiary structure make α-syn more susceptible to its surrounding environment. Under stress conditions, small organic molecules known as osmolytes accumulate inside the cells.

Bayesian Nonparametrics for FRET using Realistic Integrative Detectors.

Förster resonance energy transfer (FRET) is a widely used tool to probe nanometer scale dynamics, projecting rich 3D biomolecular motion onto noisy 1D traces. However, interpretation of FRET traces remains challenging due to degeneracy-distinct structural states map to similar FRET efficiencies- and often suffers from under- and/or over-fitting due to the need to predefine the number of FRET states and noise characteristics. Here we provide a new software, Bayesian nonparametric FRET (BNP-FRET) for binned data obtained from integrative detectors, that eliminates user-dependent parameters and accurately incorporates all known noise sources, enabling the identification of distinct configurations from 1D traces in a plug-n-play manner.

DNA origami-based platform for multi-axial single-molecule force spectroscopy reveals hidden dynamics in Holliday junctions.

In living cells, biomolecules operate in a crowded 3D milieu and are subjected to complex multi-axial stress environment. These mechanical forces are fundamental regulators of biomolecular structures and functions. However, most single-molecule force spectroscopy techniques primarily exert force along a single axis, thereby failing to recreate the mechanical environments experienced by biomolecules in cells.